Synthetic soil-extract materials and medicaments for influenza viruses based thereon

ABSTRACT

Phenolic polymers are prepared by oxidizing and polymerizing starting organic compounds comprising at least one hydroxyl group and at least one carbonyl group or at least two hydroxyl groups on an aromatic structure. One or more inorganic compounds or salts is added and the solution is allowed to stand at about 20° C. to 80° C. for a period of about at least 2 hours. Salt molecules as well as starting compounds and other low molecular-weight materials below about 500 to about 10,000 daltons are removed from the product solution. Purified phenolic polymers are prepared in concentrated aqueous solution or in dried powder form in a final step, if necessary. The resultant phenolic polymers exhibit physicochemical properties strongly resembling those of typical commercially-available natural-product soil extracts. The materials are active influenza anti-viral agents, and are effective in anti-viral compositions for treating or preventing influenza viral diseases.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit under 35 U.S.C. §119(e) of U.S.Provisional Application Ser. No. 60/297,669, filed Jun. 12, 2001 and isa continuation-in-part of application Ser. No. 09/345,865, filed Jul. 1,1999, which is a divisional of application Ser. No. 08/798,329, filedFeb. 10, 1997, now U.S. Pat. No. 5,945,446, issued Aug. 31, 1999.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to synthetic soil extract substances comprised ofphenolic polymers, and to compositions and methods for employing thesesynthetic phenolic polymers for preventing, reducing, treating, oreliminating influenza viral diseases.

2. Description of Related Art

Soil extract materials, particularly the classes of substances knowncollectively as “humus,” “humics,” “humic acid(s),” or “humates,” havebeen widely used in a number of applications for many years, as reviewedby F. J. Stevenson, Humus Chemistry. Genesis Composition Reactions; NewYork: Wiley, 1964; and, more recently, by A. Piccolo, Humic Substancesin Terrestrial Ecosystems; New York: Elsevier, 1996.

Humic substances have long been known to exhibit anti-viral properties(H. Schultz, Dtsch. Tierarztl. Wochenschr. 1962, 69, 613; 1965, 72(13),294-297; R. Klocking and M. Sprossig, Experientia 1972, 28(5), 607-608),particularly retroviruses (G. Sydow, V. Wunderlich, R. Klocking, and B.Helbig, Pharmazie 1986, 41(12), 865-868). Viral pathogens for whichsoil-extract materials have been shown to be effective include inparticular Coxsackie virus A9 (Griggs-Baylor) (R. Klocking and M.Sprossig, Experientia 1972, 28(5), 607-608), herpes simplex virus type 1(B. T. Rouse (Ed.), Herpes Simplex Virus; Berlin: Springer-Verlag, 1992;R. Klocking, K. D. Thiel, P. Wutzler, B. Helbig, and P. Drabke,Pharmazie 1978, 33(8), 539; F. Schiller, R. Klocking, P. Wutzler, and I.Farber, Dermatol. Monatsschr. 1979, 165(7), 505-509; B. Helbig, A.Sauerbrei, R. Klocking, P. Wutzler, N. Wicht, U. Wiedemann, and G.Herrmann, J. Med. Virol. 1987, 23(3), 303-309; R. Klocking and B.Helbig, in Humic Substances in the Aquatic and Terrestrial Environment;Berlin: Springer-Verlag, 1991; 407-412;) and type 2 (anon. Zentralbl.Bakteriol [Orig. A] 1976, 234(2), 159-169; K. D. Thiel, R. Klocking, H.Schweizer, and M. Sprossig, Zentralbl. Bakteriol [Orig. A] 1977, 239(3),304-321; K. D. Thiel, B. Helbig, R. Klocking, P. Wutzler, M. Sprossig,and H. Schweizer, Pharmazie 1981, 36(1), 50-53; K. D. Thiel, B. Helbig,M. Sprossig, R. Klocking, and P. Wutzler, Acta Virol. 1983, 27(3),200-208; K. D. Thiel, P. Wutzler, B. Helbig, R. Klocking, M. Sprossig,and H. Schweizer, Pharmazie 1984, 39(11), 781-782); humanimmunodeficiency virus (HIV) (M. Cushman, P. Wang, S. H. Chang, C. Wild,E. De Clercq, D. Schols, M. E. Goldman, and J. A. Bowen, J. Med. Chem.1991, 34(1), 329-337; M. Cushman, S. Kanamathareddy, E. De Clercq, D.Schols, M. E. Goldman, and J. A. Bowen, J. Med. Chem. 1991, 34(1),337-342; D. Schols, P. Wutzler, R. Klocking, B. Helbig, and E. DeClercq, J. Acquir. Immune Defic. Syndr. 1991, 4(7), 677-685; S. Loya, R.Tal, A. Hizi, S. Issacs, Y. Kashman, and Y. Loya, J. Nat. Prod. 1993,56(12), 2120-2125; J. Schneider, R. Weis, C. Manner, B, Kary, A. Werner,B. J. Seubert, and U. N. Riede, Virology 1996, 218(2), 389-395;influenza virus type A (Krasnodar/101/59/H2N2) (R. Mentel, B. Helbig, R.Klocking, L. Dohner, and M. Sprossig, Biomed. Biochim. Acta 1983,42(10), 1353-1356); and type B (J. Hils, A. May, M. Sperber, R.Klocking, B. Helbig, and M. Sprossig, Biomed. Biochim. Acta 1986, 45(9),1173-1179); as well as other respiratory tract infectious agents (A.Jankowski, B. Nienartowicz, B. Polanska, and A. Lewandowicz-Uszyuska,Arch. Immunol. Ther. Exp. (Warsz) 1993, 41(1), 95-97).

The mechanisms whereby humic substances inhibit the cytopathicity of anumber of viruses have been studied in some detail. It is thought thatthe materials prevent viral replication in part by sorbing onto theviral envelope protein (gp120 in the case of HIV), and thereby block thesorption of viral particles to cell surfaces: K. D. Thiel, R. Klocking,H. Schweizer, and M. Sprossig, Zentralbl. Bakteriol. [Orig. A] 1977,239(3), 304-321; D. Schols, P. Wutzler, R. Klocking, B. Helbig, and E.De Clercq, J. Acquir. Immune Defic. Syndr. 1991, 4(7), 677-685; anon.,Fortschr. Med. 1995, 113(7), 10; J. Schneider, R. Weis, C. Manner, B.Kary, A. Werner, B. J. Seubert, and U. N. Riede, Virology 1996, 218(2),389-395. [Extracellular interception of pathogens by chemical agentsthat bind to them is a well-known means of immunological defense (D. M.Shankel, S. Kuo, C. Haines, and L. A. Mitscher, in Antimutagenesis andAnticarcinogenesis Mechanisms III; G. Bronzetti, H. Hayatsu, S. DeFlora, M. D. Waters, and D. M. Shankel (Eds.); New York: Plenum, 1993;65-74). Such materials might well be termed “despathogens”, followingthe terminology proposed by T. Kada and K. Shimoi, Bioessays 1987, 7,113-116, regarding “desmutagens”.] It has also been found thatnaturally-occurring humic acid preparations can stimulate the productionof cytokines, including interferon-gamma, interferon-alpha, and tumornecrosis factor-alpha (A. D. Inglot, J. Zielinksa-Jenczylik, and E.Piasecki, Arch. Immunol. Ther. Exp. (Warsz) 1993, 41(1), 73-80); as wellas interferon-beta (Z. Blach-Olszewska, E. Zaczynksa, E. Broniarek, andA. D. Inglot, Arch. Immunol. Ther. Exp. (Warsz), 1993, 41(1), 81-85).

The toxicity of naturally-occurring humic acids is remarkably low (K. D.Thiel, B. Helbig, R. Klocking, P. Wutzler, M. Sprossig, and H.Schweizer, Pharmazie 1981, 36(1), 50-53; U. N. Riede, I. Jonas, B. Kim,U. H. Usener, W. Kreutz, and W. Schlickewey, Arch. Orthop. Trauma Surg.1992, 111(5), 259-264; H. Czyzewska-Szafran, Z. Jastrzebski, D.Soltysiak-Pawluczak, M. Wutkiewicz, A. Jedrych, and M. Riemiszewska,Acta Pol. Pharm. 1993, 50(4-5), 373-377; H. L. Yang, F. J. Lu, S. L.Wung, and H. C. Chiu, Thromb. Haemost. 1994, 71(3), 325-330). [Cytotoxiceffects of anti-viral substances, including humic acids, are usuallyevaluated via biological (viability and alterations of cell morphology)and biochemical testing methods (⁵¹Cr release), as described by K. D.Thiel, U. Eichhom, H. Schweizer, and R. Klocking, Arch. Toxicol. Suppl.1980, 4, 428-430.] The cytotoxicity (CD₅₀) of a naturally-occurringhumic acid for human peripheral blood leukocytes (PBL) was found to be1-9 milligrams per milliliter. In addition, J. Schneider, R. Weis, C.Manner, B. Kary, A. Werner, B. J. Seubert, and U. N. Riede, Virology1996, 218(2), 389-395, reported that the cytotoxicity of a synthetichumic acid prepared from hydroquinone for MT-2 cells was approximately600 micrograms per milliliter. It has also been found that medicamentsprepared from humic acids isolated from naturally-occurring soilmaterials are neither carcinogenic (Syrian hamster embryo celltransformation test: J. Koziorowska and E. Anuszewska, Acta Pol. Pharm.1994, 51(1), 101-102) nor mutagenic (T. Sato, Y. Ose, and H. Hagase,Mutat. Res. 1986, 162(2), 173-178; V. M. Sui, A. I. Kiung, and T. I.Veidebaum, Vopr. Kurortol. Fiozioter. Lech. Fiz. Kult. 1986, 2(3-4),34-37; J. Koziorowska, B. Chlopkiewicz, and E. Anuszewska, Acta Pol.Pharm. 1993, 50(4-5), 379-382). Prenatal (S. Golbs, V. Fuchs, M.Kuhnert, and C. Polo, Arch. Exp. Veterinarmed. 1982, 36(2), 179-185) andembryotoxic and teratogenic effects (T. Juszkiewicz, M. Minta, B.Wlodarczyk, B. Biernacki, and J. Zmudzki, Acta Pol. Pharm. 1993,50(4-5),383-388) are also not observed with humic preparations at dailydose levels from 5-50 milligrams per kilogram body weight. Topicalpreparations are tolerated even better (V. V. Soldatov and M. N.Cherepanova, Vopr. Kurortol. Fizioter. Lech. Fiz. Kult. 1970, 35(3),256-259; H. Czyzewska-Szafran, Z. Jastrzebski, D. Soltysiak-Pawluczuk,M. Wutkiewicz, A. Jedrych, and M. Remiszewska, Acta Pol. Pharm. 1993,50(4-5), 373-377) when applied dermally in aqueous solution in amountsas high as 10 percent weight-by-volume (K. Wiegleb, N. Lange, and M.Kuhnert, Dtsch. Tierarztl. Wochenschr. 1993, 100(10), 412-416).

Because humic substances are not chemically well-defined, thepreparation of synthetic humic acids whose physicochemical propertiesmimic naturally-occurring materials is quite difficult, as pointed outby K. Murray and P. W. Linder, J. Soil Sci. 1983, 34, 511-523.Nevertheless, there have been several notable advances in this area.Broadly speaking, three general strategies have evolved. All depend uponstarting with well-defined molecules of molecular weight on the order ofhydroxybenzoic acid, and then causing the molecules to polymerize uponthemselves to form larger molecules. The methods differ in the causationfactor, which can be microbial, chemical, or enzymatic.

Humic acids of microbial origin have been described and discussed by M.Robert-Gero, C. Hardisson, L. Le Borgne, and G. Pignaud, Ann. Inst.Pasteur (Paris) 1966, 111(6), 750-767; and by M. Robert-Gero, C.Hardisson, L. Le Borgne, and G. Vidal, Ann. Inst. Pasteur (Paris) 1967,113(6), 903-909.

The chemical synthesis of humic acids has been pioneered by R. Klocking,B. Helbig, and associates: R. Klocking, B. Helbig, and P. Drabke,Pharmazie 1977, 32, 297; R. Klocking, B. Helbig, K. D. Thiel, T.Blumohr, P. Wutzler, M. Sprossig, and F. Schiller, Pharmazie 1979,34(5-6), 293-294; R. Mentel, B. Helbig, R. Klocking, L. Dohner and M.Sprossig, Biomed. Biochim. Acta 1983, 42(10), 1353-1356; H. P. Klocking,R. Klocking, and B. Helbig, Farmakol. Toksikol. 1984, 47(1), 93-95; K.D. Thiel, P. Wutzler, B. Helbig, R. Klocking, M. Sprossig, and H.Schweizer, Pharmazie 1984, 39(11), 781-782; J. Hils, A. May, M. Sperber,R. Klocking, B. Helbig, and M. Sprossig, Biomed. Biochim. Acta 1986,45(9), 1173-1179; B. Helbig, A. Sauerbrei, R. Klocking, P. Wutzler, N.Wicht, U. Wiedemann, and G. Herrmann, J. Med. Virol. 1987, 23(3),303-309; K. I. Hanninen, R. Klocking, and B. Helbig, Sci. Total Environ.1987, 62, 201-210; R. Klocking and B. Helbig, in Humic Substances in theAquatic and Terrestrial Environment; New York: Springer-Verlag, 1989;407-412; C. Schewe, R. Klocking, B. Helbig, and T. Schewe, Biomed.Biochim. Acta 1991, 50(3), 299-305; D. Schols, P. Wutzler, R. Klocking,B. Helbig, and E. De Clercq, J. Acquir. Immune Defic. Syndr. 1991, 4(7),677-685. Typically, 10 millimoles of the starting small-moleculephenolic compound is dissolved in distilled water, the pH is adjusted to8.5 with aqueous sodium hydroxide (NaOH), and then 2-5 millimoles ofsodium periodate (NaIO₄) is added. The solution is warmed at 50° C. for30 minutes, and is then allowed to stand overnight. The resultant humicacid-like polymeric products are isolated by precipitation with lead(II)nitrate [Pb(NO₃)₂]. The precipitated polymers are redissolved in aqueoussodium hydroxide (pH 8.5) and heated with 8-hydroxyquinoline for 30minutes at 100° C. The precipitate formed is lead(II) chelate, which isremoved by filtration. Residual 8-hydroxyquinoline is extracted withchloroform, and the desired polymeric material is then precipitated fromthe aqueous solution by the addition of various combinations of aceticacid, ethyl acetate, and ethanol. Starting compounds that have been usedfor the synthesis of humic-like materials include4-[bis(p-hydroxyphenyl)methylene]-2,5-cyclohexadien-1-one (aurin),4-[bis(3-carboxy-4-hydroxyphenyl)methylene]-2-carboxy-2,5-cyclohexa-dien-2-one(aurintricarboxylic acid), 3-(3,4-dihydroxyphenyl)propenoic acid(caffieic acid), 1,2-dihydroxybenzene (catechol),1,3,4,5-tetrahydroxycyclohexanecarboxylic acid3-(3,4-dihydroxyphenyl)propenoate (chlorogenic acid),3,4-dihydroxyphenylacetic acid (homoprotocatechuic acid),1-(3,4-dihydroxyphenyl)-2-(N-methylamino)ethanol (epinephrine),3-(4-hydroxy-3-methoxyphenyl)-2-propenoic acid (ferulic acid),3,4-5-trihydroxybenzoic acid (gallic acid), 2,5-dihydroxybenzoic acid(gentisic acid), 2,5-dihydroxyphenylacetic acid (homogentisic acid),3-(3,4-dihydroxyphenyl)propionic acid (hydrocaffeic acid),1,4-dihydroxybenzene (hydroquinone), 2,3-dihydroxytoluene(3-methylcatechol), 3,4-dihydroxytoluene (4-methylcatechol),2,5-dihydroxytoluene (2-methylhydroquinone),4,4′-(2,3-dimethyltetramethylene)-di-(1,2-dihydroxybenzene)(nordihydroguaiaretic acid), 1-(3,4-dihydroxyphenyl)-2-aminoethanol(norepineph-rine), 3,4-dihydroxybenzoic acid (protocatechuic acid),1,2,3-trihydroxybenzene (pyrogallol), 1,3-dihydroxybenzene (resorcinol),and 4-hydroxy-3-methoxybenzoic acid (vanillic acid). Other notableefforts on the chemical synthesis of humic-like substances include thestudies by De Clercq and colleagues on aurintricarboxylic acid, itsderivatives, and related compounds: M. Cushman, P. Wang, S. H. Chang, C.Wild, E. De Clercq, D. Schols, M. E. Goldman, and J. A. Bowen, J. Med.Chem. 1991, 34(1), 329-337; M. Cushman, S. Kanamathareddy, E. De Clercq,D. Schols, M. E. Goldman, and J. A. Bowen, J. Med. Chem. 1991, 34(1),337-342. Related efforts have also been reported by M. Robert-Gero, C.Hardisson, L. Le Borgne, and G. Vidal, Ann. Inst. Pasteur (Paris) 1967,113(6), 903-909; M. Jakubiec, E. Miszczak, and J. Szczerkowska, ActaMicrobiol. Pol. [B] 1971, 3(1), 63-66; R. Ansorg and W. Rochus,Arzneimittelforschung 1978, 28(12), 2195-2198; J. Pommery, M. Imbenotte,A. F. Urien, D. Marzin, and F. Erb, Mutat. Res. 1989, 223(2), 183-189;F. J. Lu and Y. S. Lee, Sci. Total Environ. 1992, 114, 135-139; K.Wiegleb, N. Lange, and M. Kuhnert, DTW Dtsch. Tierarztl. Wochenschr.1993, 100(10), 412-416; H. L. Yang, F. J. Lu, S. L. Wung, and H. C.Chiu, Thromb. Haemost. 1994, 71(3), 325-330; W. Seffner, F. Schiller, R.Heinze, and R. Breng, Exp. Toxicol. Pathol. 1995, 47(1), 63-70; and J.Schneider, R. Weis, C. Manner, B. Kary, A. Werner, B. J. Seubert, and U.N. Riede, Virology 1996, 218(2), 389-395.

The enzymatic catalytic synthesis of humic acids dates to about 1961with the work by R. E. Hampton and R. W. Fulton, Virology 1961, 13,44-52 (see also R. E. Hampton, Phytophathology 1970, 60, 1677-1681), whofound that enzymatically oxidized phenols inactivate phytopathogenic(i.e., plant-related) viruses. Typically o-diphenol oxidase has beenemployed for the enzymatic synthesis of humic-like materials: anon.Zentralbl. Bakteriol. [Orig A] 1976, 234(2), 159-169; R. Klocking, B.Helbig, and P. Drabke, Pharmazie 1977, 32(5), 297; K. D. Thiel, B.Helbig, R. Klocking, P. Wutzler, M. Sprossig, and H. Schweizer,Pharmazie 1981, 36(1), 50-53; K. D. Thiel, B. Helbig, M. Sprossig, R.Klocking, and P. Wutzler, Acta Virol. 1983, 27(3), 200-208; K. D. Thiel,P. Wutzler, B. Helbig, R. Klocking, M. Sprossig, and H. Schweizer,Pharmazie 1984, 39(11), 781-782; and G. Sydow, V. Wunderlich, R.Klocking, and B. Helbig, Pharmazie 1986, 41(12), 865-868.

A direct comparison of humic acids synthesized enzymatically andnonenzymatically from caffeic and hydrocaffeic acids has shown that thetwo synthetic routes produce materials that differ somewhat in theirefficacy for the suppression of herpes (hominis) types 1 and 2 viruses:K. D. Thiel, P. Wutzler, B. Helbig, R. Klocking, M. Sprossig, and H.Schweizer, Pharmazie 1984, 39(11), 781-782.

PCT application WO 00/16785 (Mar. 20, 2000) from Dekker and Medlendiscloses the use of humic acid or its salts, esters, or derivativesthereof, all prepared as described in U.S. Pat. Nos. 4,912,256 and5,004,831 from coal extracts, in stimulating lymphocytes in a human,animal, or bird. This allows for the treatment of viral and bacterialinfections, and more particularly HIV infections, cancer, andopportunistic diseases. Oxihumic acids, salts, esters, or derivativesthereof are preferred. Administration is preferably oral. Some examplepharmacological data presented include the antiviral activity ofoxihumates against HIV-1 in vitro and clinical trials of oral oxihumatein HIV-infected patients.

PCT application WO 00/16786 (Mar. 30, 2000) from Dekker and Medlendiscloses the use of pharmaceutical compositions comprising an oxihumicacid or its salts, esters, or derivatives thereof, all prepared asdescribed in U.S. Pat. Nos. 4,912,256 and 5,004,831 from coal extracts,as active ingredients. Compositions are preferably administered orallyfor stimulating lymphocytes in a human, animal, or bird. They may beused in treating viral and bacterial infections, HIV infections,opportunistic diseases, inflammation, pain and fever, cancer growth, anddiseases associated with viral infection and a depressed immune system.A number of pharmacological examples are given, including interleukin 10production by oxihumate-reated lymphocytes, increased antibodyproduction against Newcastle disease in chickens treated with oxihumate,TNF production by oxihumate-treated lymphocytes, and antiviral activityof oxihumate against HSV-1 and coxsackie virus type 1 in vitro.

The diversity of physicochemical characteristics as well as widevariation in the biological activity and toxicity of humics extracted orotherwise derived from natural soils has been well documented. Thisdiversity and variation is due to variations in factors such as thesource of the soil, the method(s) of extraction and/or isolation, andthe technique(s) employed to treat the extract once it has beenseparated and isolated from crude soil. The consequence ofirreproducibility of the properties of substances extracted from naturalsoil is that the commercial value of such materials is minimized. Inaddition, they are rendered unsuitable as medicaments. Also, while anumber of laboratory-scale processes have already been described thataddress various aspects of the isolation, synthesis, and/or preparationof humic substances or similar materials, there are no reports ofpreparing and isolating such purely synthetic humic acids or similarmaterials by methods that are suitable for scaleup directly toindustrial levels, that provide economically acceptable yields, and thatoptimize the preparation procedures from the standpoint of medicamentsafety and efficacy. All of the known synthetic methods utilizepotentially toxic precipitation methods [lead(II) nitrate precipitation]followed by complex isolation procedures, potentially mutageniccompound-producing hydrochloric acid precipitation or lengthy syntheticsteps as long as 10 days.

SUMMARY OF THE INVENTION

There is a need to devise simple synthetic procedures that yieldinexpensive, safe materials whose physicochemical attributes arereproducible, and that at least simulate those of typicalcommercially-available soil extracts.

One embodiment is a method for preventing and/or treating influenzavirus infection in a mammal which comprises administering an effectiveamount of a synthetic phenolic polymeric material which is prepared by:

a) dissolving in an aqueous solution at least one starting organiccompound comprising at least one hydroxyl group and at least onecarbonyl group or at least two hydroxyl groups on an aromatic structure;

b) adjusting the pH of the aqueous solution resulting from step a) tobetween about 8 and 11;

c) adding an alkaline periodate salt or alkaline-earth periodate salt tothe aqueous solution resulting from step b);

d) maintaining the temperature of the solution from step c) betweenabout 20° C. and 100° C. for a period of at least about 30 minutes;

e) adding at least one water soluble compound or salt selected from thegroup consisting of boric acid, borate salts, alkaline earth salts,transition metal salts, alkaline sulfides, alkaline earth sulfides, ortransition metal sulfides to the aqueous solution resulting from stepd);

f) allowing the aqueous solution from step e) to stand with or withoutstirring at about 20° C. to 100° C. for at least about 2 hours; and

g) removing molecules from the solution resulting from step f) belowabout 500 to about 10,000 daltons.

In another aspect, the method of preparation of the synthetic phenolicpolymeric material further comprises a step, following the step ofremoving molecules from the solution below about 500 daltons to 10,000daltons, of concentrating the solution.

In another aspect, the method of preparation of the synthetic phenolicpolymeric material further comprises a step, following the step ofremoving molecules from the solution below about 500 dalton to 10,000daltons, of removing water from the solution.

In another aspect, the influenza virus infection is effected by a virus,preferably influenza A or influenza B.

In another aspect, the administering of a synthetic phenolic polymericmaterial is along with an effective amount of an antiviral composition.

In another aspect, the administering of a synthetic phenolic polymericmaterial can be achieved systemically or topically.

One embodiment is a method for inhibiting influenza viral attachment tohost cells in a mammal which comprises administering an effective amountof a synthetic phenolic polymeric material which is prepared by:

a) dissolving in an aqueous solution at least one starting organiccompound comprising at least one hydroxyl group and at least onecarbonyl group or at least two hydroxyl groups on an aromatic structure;

b) adjusting the pH of the aqueous solution resulting from step a) tobetween about 8 and 11;

c) adding an alkaline periodate salt or alkaline-earth periodate salt tothe aqueous solution resulting from step b);

d) maintaining the temperature of the solution from step c) betweenabout 20° C. and 100° C. for a period of at least about 30 minutes;

e) adding at least one water soluble compound or salt selected from thegroup consisting of boric acid, borate salts, alkaline earth salts,transition metal salts, alkaline sulfides, alkaline earth sulfides, ortransition metal sulfides to the aqueous solution resulting from stepd);

f) allowing the aqueous solution from step e) to stand with or withoutstirring at about 20° C. to 100° C. for at least about 2 hours; and

g) removing molecules from the solution resulting from step f) belowabout 500 to about 10,000 daltons.

In another aspect, the method of preparation of the synthetic phenolicpolymeric material further comprises a step, following the step ofremoving molecules from the solution below about 500 daltons to 10,000daltons, of concentrating the solution.

In another aspect, the method of preparation of the synthetic phenolicpolymeric material further comprises a step, following the step ofremoving molecules from the solution below about 500 dalton to 10,000daltons, of removing water from the solution.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a high-performance liquid chromatography (HPLC) trace forthe synthetic humic acid product obtained from 2,5-dihydroxyphenylaceticacid (homogentisic acid), as described in Examples 10, 11, and 14.

FIG. 2 shows a high-performance liquid chromatography (HPLC) traceobtained for a typical commercially-available natural-product humicacid.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

U.S. Pat. No. 5,945,446, issued Aug. 31, 1999, discloses the process forpreparing synthetic soil-extract materials and medicaments basedthereon. U.S. Pat. No. 5,945,446 and the references therein areincorporated herein by reference in their entirety.

The inventor has developed combinations of chemical processes for thepreparation of synthetic phenolic polymeric materials, also known assynthetic humic acids, whose physicochemical properties and attributesare reproducible, and which simulate those of typicalcommercially-available natural humic acids and other soil extracts,which contain little or no ionic salts or other compounds of molecularweight less than about 500 daltons, which have a minimum molecularweight of about 500 daltons, and which processes shall be suitable forscaleup directly to industrial levels that provide economicallyacceptable yields.

The inventor has also developed compositions and methods for treating orpreventing influenza viral diseases by using an effective amount foranti-viral activity of a synthetic humic acid prepared according to theabove processes.

The starting compounds used in the chemical processes employed forproduction of synthetic humic acids according to embodiments of themethod of the preferred embodiments are known materials that are readilyavailable commercially.

Generally speaking, the chemical processes for the preparation ofsynthetic humic acids of the preferred embodiments include the followingsteps:

a) dissolving in an aqueous solution at least one starting organiccompound comprising at least one hydroxyl group and at least onecarbonyl group or at least two hydroxyl groups on an aromatic structure;

b) adjusting the pH of the aqueous solution resulting from step a) tobetween about 8 and 11;

c) adding an alkaline periodate salt or alkaline-earth periodate salt tothe aqueous solution resulting from step b);

d) maintaining the temperature of the solution from step c) betweenabout 20° C. and 100° C. for a period of at least about 30 minutes;

e) adding at least one water soluble compound or salt selected from thegroup consisting of boric acid, borate salts, alkaline earth salts,transition metal salts, alkaline sulfides, alkaline earth sulfides, ortransition metal sulfides to the aqueous solution resulting from stepd);

f) allowing the aqueous solution from step e) to stand with or withoutstirring at about 20° C. to 100° C. for at least about 2 hours; and

g) removing molecules from the solution resulting from step f) belowabout 500 to about 10,000 daltons;

h) concentrating the solution resulting from step g), if necessary; and

i) removing the water from the solution resulting from step h), ifnecessary.

The starting organic compound in step a) above can be one, or more thanone in combination, of different compounds taken from the groupconsisting of starting organic compounds illustrated in Tables 1 and 2.Starting organic compounds illustrated in Table 1 are comprised of asingle benzene ring with six substituents R₁-R₆, wherein R₁-R₆ can beany one of the indicated atom or functional groups, as long as at leastone of R₁-R₆ is a hydroxy (—OH) functional group. Preferably, at leastone of R₁-R₆ is a hydroxy (—OH) functional group and at least one of theremaining substituents R₁-R₆ contains a carboxylic acid functionalgroup. More preferably, two of R₁-R₆ are hydroxy (—OH) functional groupsand one of the remaining substituents R₁-R₆ contains a carboxylic acidfunctional group. Homogentisic acid, which is 2,5-dihydroxyphenylaceticacid, is a particularly preferred starting organic compound. Caffeicacid, which is 3,4-dihydroxycinnamic acid, is another particularlypreferred starting organic compound. Chlorogenic acid, which is1,3,4,5-tetrahydroxycyclohexanecarboxylic acid3-(3,4-dihydroxycinnamate) is yet another particularly preferredstarting organic compound.

TABLE 1

R₁, R₂, R₃, R₄, R₅, R₆ = —H —CH₃ —CH₂CH₃ —(CH₂)₂CH₃ —CH(CH₃)₂ —OH —OCH₃—CHO —CO₂H —CO₂CH₃ —CH₂OH —CH₂OCH₃ —CH₂CHO —CH₂CO₂H —CH₂CO₂CH₃ —(CH₂)₂OH—(CH₂)₂OCH₃ —(CH₂)₂CHO —(CH₂)₂CO₂H —(CH₂)₂CO₂CH₃ —CH(CH₃)OH —CH(CH₃)OCH₃—CH(CH₃)CHO —CH(CH₃)CO₂H —CH(CH₃)CO₂CH₃ —CH(CH₃)CH₂OH —CH(CH₃)CH₂OCH₃—CH(CH₃)CH₂CHO —CH(CH₃)CH₂CO₂H —CH(CH₃)CH₂CO₂CH₃ —CH(OH)₂ —CH(OH)OCH₃—CH(OH)CHO —CH(OH)CO₂H —CH(OH)CO₂CH₃ —CH(OCH₃)OH —CH(OCH₃)₂ —CH(OCH₃)CHO—CH(OCH₃)CO₂H —CH(OCH₃)CO₂CH₃ —CH(OH)CH₂OH —CH(OH)CH₂OCH₃ —CH(OH)CH₂CHO—CH(OH)CH₂CO₂H —CH(OH)CH₂CO₂CH₃ —CH(OCH₃)CH₂OH —CH(OCH₃)CH₂OCH₃—CH(OCH₃)CH₂CHO —CH(OCH₃)CH₂CO₂H —CH(OCH₃)CH₂CO₂CH₃ —(CH₂)₃OH—(CH₂)₃OCH₃ —(CH₂)₃CHO —(CH₂)₃CO₂H —(CH₂)₃CO₂CH₃ —CHCHOH (cis or trans)—CHCHOCH₃ (cis or trans) —CHCHCHO (cis or trans) —CHCHCO₂H (cis ortrans) —CHCHCO₂CH₃ (cis or trans) —CH₂CHCHOH (cis or trans) —CH₂CHCHOCH₃(cis or trans) —CH₂CHCHCHO (cis or trans) —CH₂CHCHCO₂H (cis or trans)—CH₂CHCHCO₂CH₃ (cis or trans)

TABLE 2

Nordihydroguaiaretic Acid Chlorogenic Acid

Epinephrine Norepinephrine

Aurin Aurintricarboxylic Acid

Tetrahydroxybenzoquinone

Various initial concentrations of starting organic compounds in water,preferably distilled, can be employed and no lower or upper limits areuniformly required. A low concentration solution of sodium hydroxide,such as about 0.1 Normal, may also be employed as a diluent for thestarting organic compound. The appropriate initial concentration of thestarting organic compound or compounds is determined by the synthesisyield requirements and inherent requirements, such as the upper limit ofaqueous solubility of the starting organic compound or compounds.Conventional methods are employed to determine the appropriate initialconcentration of the starting organic compound or compounds.

The pH of the aqueous solution containing the starting organic compoundor compounds can be adjusted in step b) to between about 8 and 11 byadding aqueous ammonium hydroxide, or other aqueous alkaline oxide orhydroxide, or aqueous alkaline earth oxide or hydroxide, or aqueoustransition metal oxide or hydroxide. Additionally, if the initialaqueous solution contains a low concentration of base, such as about 0.1Normal sodium hydroxide and the initial solution pH is too high, an acidsuch as hydrochloric acid may be employed to adjust the pH to thedesired value. Other inorganic acids may also be employed for pHadjustment. Note that if hydrochloric acid is employed to adjust the pHdownwards from an initial high value, care should be taken to avoidletting the pH go below about 8. Acidic conditions below about pH 7should be avoided in the presence of hydrochloric acid to eliminate thepossibility of formation of mutagenic chlorinated humic acid materials.

An alkaline periodate salt or alkaline earth periodate salt may beemployed as an oxidant or polymerization initiator of the startingorganic compound in step c). Sodium periodate is particularly preferred.The concentration of the alkaline periodate salt or alkaline earthperiodate salt is generally between about 10% and 100% of the startingorganic compound or compounds on a molar basis. Thus, if 10 millimolesof starting organic compound is employed, 1 to 10 millimoles of alkalineperiodate salt may be employed. Preferably, a molar concentration ofperiodate that is about 10%-50% of the molar concentration of thestarting organic compound or compounds is employed. Most preferably, amolar concentration of periodate that is about 25%-35% of the molarconcentration of the starting organic compound or compounds is employed.The exact concentration to be used can be determined by conventionalsynthetic yield optimization techniques.

Alkaline or alkaline earth sulfides or transition metal sulfides can beoptionally added to the initial aqueous solution containing the startingorganic compound or compounds following the pH adjustment in step b) andimmediately before, at the same time, or following the addition of theperiodate in step c). Sulfides contribute to the phenolic polymericstructure, the stability of the structure and its biological activity.Sodium sulfide nonahydrate is a particularly preferred sulfide. Theconcentration of the sulfide is generally between about 1% and 20% ofthe starting organic compound or compounds on a molar basis. Thus, if 10millimoles of starting organic compound is employed, 0.1 to 2 millimolesof sulfide may be employed. Preferably, a molar concentration of sulfidethat is about 5%-15% of the molar concentration of the starting organiccompound or compounds is employed. Most preferably, a molarconcentration of sulfide that is about 8% to 12% of the molarconcentration of the starting organic compound or compounds is employed.The exact concentration of sulfide to be used can be determined byconventional synthetic yield optimization techniques.

Steps b) and c) above give conditions for oxidizing and polymerizing thestarting organic compound. Although the use of periodate salt in basicconditions is preferable, there are other conditions that can performoxidation and polymerization of the starting organic compound. One maysubstitute other reagents known in the art that are known to performthis function. If the reagents for oxidation and polymerization aresubstituted, the temperature and time period for the reaction in step d)should be adjusted accordingly for optimization. For example, a phenolicsolution with about 2 equivalents of hydrogen peroxide can react forabout one week at about 23° C. to form humic acids.

The pH-adjusted aqueous solution containing the starting organiccompound, periodate and optional sulfide is placed in a water-bath orother thermostat heating device at about 20° C. to 100° C. for a periodof about 30 minutes to 100 hours in step d). Alternatively, the aqueoussolution itself may be thermostated between about 20° C. and 100° C. fora period of about 30 minutes to 100 hours. A preferred temperature andtime period is between about 35° C. and 80° C. for about 30 minutes to100 hours. A particularly preferred temperature and time is about 50° C.for about 30 minutes to two hours. Alternative temperatures andpressures that are equivalent to the above temperature and pressures maybe used.

Following this period, salts are added to the solution resulting fromstep d) alone or in combination in step e). Salts containing boron,calcium and other alkaline earths, iron and other transition metals arepreferred. Such salts may contribute to the phenolic polymericstructure, its stability and biological activity. Boric acid orboron-containing-borate salts, such as sodium borate, are particularlypreferred, as are alkaline earth salts, such as calcium sulfatedihydrate, and transition metal salts, such as ferrous sulfateheptahydrate. The concentrations of each of the salts employed isgenerally between about 0.1% and 20% of the starting organic compound orcompounds on a molar basis. Preferably, a molar concentration of saltwhich is about 0.2% to 10% of the molar concentration of the startingorganic compound or compounds is employed. Most preferably, a molarconcentration of salt that is about 0.2% to 2% of the molarconcentration of the starting organic compound or compounds, isemployed. The exact concentration to be used can be determined byconventional synthetic yield optimization techniques. The solutionresulting from step e) is allowed to stand at between about 20° C. and100° C. with or without stirring for a period of at least 2 hours instep f). Preferably, the solution is allowed to stand at between about20° C. and 80° C. for about 2 to 48 hours. Alternative temperatures andpressures that are equivalent to the above temperature and pressures maybe used. Any precipitate formed at this stage is removed viaconventional centrifugation.

Molecules below about 500 to about 10,000 daltons in the solutionresulting from step f) are removed in step g). A variety of knownconventional techniques can be employed, such as preparativechromatography, ultrafiltration or dialysis. Molecules are preferablyremoved from the solution resulting from step f) by employing dialysisin step g) with a flow-through open-channel or screen membrane apparatusconsisting of a sandwich-type membrane of lower molecular-weight cutoffof 500-10,000 daltons until the conductivity of the solution has droppedto about 200 microsiemens or less. Most preferably, molecules areremoved from the solution resulting from step f) by employing dialysisin step g) until the conductivity of the solution has dropped to about50 microsiemens or less. A Pall Filtron Ultrasette® Tangential FlowDevice or Mini-Ultrasette® Tangential Flow Device used with a PallFiltron Ultralab® Specialized Pump and Reservoir System are preferredfor solution dialysis.

The conductivity of the solution processed in step g) above canconveniently be monitored with a flow-through conductivity cell andconductivity meter. Alternatively, a simple inexpensive hand-heldcombination conductivity cell/conductivity meter (e.g., a NalcometerModel MLN) can be employed.

Optionally, the solution from step g) above can be concentrated. Onemethod of concentrating the solution is by removing water from thesolution. The removal of water can be accomplished by conventional meansknown in the art.

Before removing the water from the solution in step h) above, thesolution resulting from step g) above can be further dialyzed with aflow-through apparatus consisting of a sandwich-type membrane ofmolecular weight cutoff of 50,000 daltons. In this case the filtratesolution, not the retentate, is saved for further concentrating andprocessing according to steps h) and i). The resultant product will havea molecular-weight range of about 500-50,000 daltons.

If the solution resulting from either steps g) or h) above is to bestored as an aqueous solution for long periods of time for laterapplication or use, for example as an anti-viral treatment solution,anti-viral therapy, anti-microbial therapy, a spray-on fertilizer orsoil amendment, it can be filtered through standard 0.2-0.4 micronfilters to remove bacteria and viruses, that is, can be made sterile byfiltration. Alternatively, the aqueous solution from either steps g) orh) can be autoclaved for about 5-60 minutes at about 100-150° C. toproduce a sterile solution.

A final optional step i) in the process of the preferred embodimentsinvolves removing water from the solution resulting from step h). Whenfreeze-drying is employed as the method of water removal in step i)above, the resultant product is a light fluffy dark-colored powder thatis subject to static electricity effects. To minimize these effects, asmall amount of mannose or other sugar can be added to the solutionresulting from step h) just prior to freeze-drying. Water removal fromthe product can be carried out by means other than freeze-drying in stepi) above, such as by heat evaporation with or without vacuum, by rotaryevaporation, by spray-drying, or by any other solvent-removal techniquethat is convenient as well as economical for aqueous solutions. Thedried powder obtained from step i) above can be autoclaved for about15-30 minutes at about 100-120° C. to produce a sterile powder.

The synthetic humic acid materials produced according to the chemicalprocesses and separation and isolation procedures of the preferredembodiments exhibit the physicochemical properties and attributes oftypical naturally-occurring commercially-available humic acids and othersoil extracts.

A facile method of examining the physicochemical characteristics of theproduct yielded by steps a) through h) above, or by modificationsthereto, is high performance liquid chromatography (HPLC). Thechromatographic fingerprint pattern so obtained from HPLC also offers aconvenient means of comparing one product with another, as well ascomparing each of the synthetic products with naturally-occurring humicacids and other soil-extract materials. The HPLC method is thus used todetermine the reproducibility of the physiochemical properties andattributes of the synthetic phenolic polymeric materials, as well as todetermine if the aforementioned properties and attributes simulate thephysiochemical properties and attributes of typical commercial-availablenatural humic acids and other soil extracts. The latter determination ofsimulation is done in the conventional manner employing HPLC; e.g., byvisually and quantitatively comparing the HPLC chromatographicfingerprint patterns of the materials. The fingerprint patterns of thetwo materials, one synthetic and one natural, need not be 100% identicalto conclude that the physiochemical properties and attributes of thesynthetic phenolic polymeric material simulates the physiochemicalproperties and attributes of the natural humic acid. An approximatecorrespondence between the aforementioned HPLC fingerprint patterns isall that is required to conclude that the synthetic material simulatesthe natural material. In general, even about 75% visual correspondencein two HPLC fingerprint patterns is all that is necessary to concludethat one material simulates another.

A useful fingerprint pattern for natural as well as synthetic soilextract materials can be obtained as follows. The column is comprised ofa packing, typically reversed-phase polymer PRP-1 (Hamilton Co.), ofparticle size 5 microns, and being 150 millimeters in length by 4.1millimeters inside diameter. The mobile phase is comprised of threesolutions. Solution A is about 0.1 Normal aqueous sodium hydroxide.Solution B is about 0.05 Normal of so-called Prideaux universal buffer,which is made by combining about 4.25 grams of sodium nitrate (NaNO₃),about 12.37 grams of boric acid (H₃BO₃), about 23.06 grams of phosphoricacid (H₃PO₄), and about 12.01 grams of acetic acid (CH₃CO₂H) with about4 liters of distilled water. Solution C is about 100% methanol (CH₃OH).The mobile-phase gradient employed for an HPLC run consists of about 40%solution A plus about 60% solution B at the beginning, which compositionis changed in a linear manner to about 100% solution A after about 20minutes. The mobile phase is then changed linearly again to about 10% Aplus about 90% C over the next about 5 minutes, which final compositionis held for the purpose of a column wash for the next about 35 minutes.The mobile-phase flow rate is about 1 milliliter per minute. Thedetector is UV-Visible, which is set at 340 nanometers. The chart speedis typically 0.5 centimeter per minute. The sample loop size is about5-20 microliters. Solutions are prepared for HPLC by dissolving about0.1-10 grams of dried sample in about 100 milliliters of distilled wateror about 0.1 Normal aqueous sodium hydroxide of about pH 8-10.

The chemical processes and separation and isolation procedures of thepresent invention are suitable for scaleup directly to industrial levelsthat provide economically acceptable yields. The chemical processes andseparation and isolation procedures of the present invention can producesynthetic product yields approaching 100%. More typically, approximately0.08 to 0.65 g of synthetic humic acid can be produced from about 10millimoles of starting organic compound or compounds in about 300milliliters. These procedures can be scaled up to pharmaceuticalproduction scales employing about 10,000 to 20,000 liters or more ofinitial solution containing the starting organic compound or compounds.A total yield between approximately 2.7 and 21.7 kilograms of synthetichumic acid can be achieved utilizing a 10,000-liter thermally-jacketedstainless-steel tank and a concentration of starting organic compound ofabout 10 millimoles per about 300 milliliters. A single anti-viraltreatment may employ milligram amounts of synthetic humic acid. Twentykilograms of synthetic humic acid represents about 2 million units ofanti-viral product at about 10 milligrams per unit. Even at a treatmentcost of about $0.10 per unit, this amount represents about $200,000.00of synthetic humic acid. Since the starting organic compounds utilizedin the preferred embodiments are relatively inexpensive, the synthesisyields of the chemical processes and separation and isolation proceduresof the preferred embodiments are economically very acceptable.

Examples 1 through 9 are illustrative of the variety of starting organiccompounds that can be employed in the process of the preferredembodiments. It was not considered necessary to carry out all steps ofthe process of the preferred embodiments to illustrate starting compoundvariety. More particularly, Examples 1 through 9 are illustrative of allsteps of the process of the preferred embodiments with the exception ofstep e), the addition of salts.

EXAMPLE 1 Preparation of a Synthetic Humic Acid from2,5-dihydroxybenzoic Acid (Gentisic Acid)

The starting organic compound is shown in Table 1, and represented byR₁=—CO₂H; R₂, R₅=—OH; and R₃, R₄, R₆=—H. Ten millimoles (1.55 grams) ofgentisic acid was dissolved in 300 milliliters of 0.1 Normal aqueoussodium hydroxide (NaOH). The solution pH was adjusted to 8.5 with 6Normal HCl. Two and one-half millimoles (0.54 gram) of sodium periodate(NaIO₄) was added, and the solution was placed in a water-bath at 50° C.for 30 minutes. The solution was allowed to stand at room temperatureovernight. Any precipitate was removed by centrifugation. The solutionwas dialyzed with a 1,000-dalton cut-off flow-through open-channel orscreen membrane system (Pall Filtron: Ultrasette® 7 Tangential FlowDevice or Mini-Ultrasette® 7 Tangential Flow Device used with a PallFiltron Ultralab® 7 Specialized Pump and Reservoir System) to aconductivity of 30 microsiemens or less against distilled water. Thedialysis apparatus was then used to concentrate the solution to about200 milliliters. The solution can be saved at this point for further useas an aqueous solution; or it can be freeze-dried to a powder. (Five totwenty hundredths of a gram of mannose or other suitable carbohydratecan be added to the solution prior to freeze-drying to reduce staticelectricity effects associated with the freeze-dried powder.) The yieldof synthetic soil extract was 0.2 gram.

The following Examples 2-9 employ the synthesis procedure of Example 1beginning with the adjustment of solution pH.

EXAMPLE 2 Preparation of a Synthetic Humic Acid from3,4-dihydroxyphenylacetic Acid (Homoprotocatechuic Acid)

The starting organic compound, 3,4-dihydroxy-phenylacetic acid, is shownin Table 1, and represented by R₁=—CH₂CO₂H; R₃, R₄=—OH; and R₂, R₅,R₆=—H. Ten millimoles (1.68 grams) of homoprotocatechuic acid wasdissolved in 300 milliliters of 0.1 Normal aqueous sodium hydroxide(NaOH). The remaining procedure followed that of Example 1. The yield ofsynthetic soil extract was 0.24 gram.

EXAMPLE 3 Preparation of a Synthetic Humic Acid fromdl-(3,4-dihydroxyphenyl)hydroxyacetic Acid (dl-3,4-dihydroxymandelicAcid)

The starting organic compound, dl-(3,4-dihydroxyphenyl)hydroxyaceticacid, is shown in Table 1, and represented by R₁=—CH(OH)CO₂H; R₃,R₄=—OH; and R₂, R₅, R₆=—H. Ten millimoles (1.84 grams) ofdl-3,4-dihydroxymandelic acid was dissolved in 300 milliliters of 0.1Normal aqueous sodium hydroxide (NaOH). The remaining procedure followedthat of Example 1. The yield of synthetic soil extract was 0.08 gram.

EXAMPLE 4 Preparation of a Synthetic Humic Acid from AurintricarboxylicAcid

The chemical structure of the starting organic compound is shown inTable 2. Ten millimoles (4.2 grams) of aurintricarboxylic acid wasdissolved in 300 milliliters of 0.1 Normal aqueous sodium hydroxide(NaOH). The remaining procedure followed that of Example 1. The yield ofsynthetic soil extract was 4.7 grams.

EXAMPLE 5 Preparation of a Synthetic Humic Acid from3-(3,4-dihydroxyphenyl)propenoic Acid (Caffeic Acid)

The starting organic compound is shown in Table 1, and represented byR₁=—CHCHCO₂H; R₃, R₄=—OH; and R₂, R₅, R₆=—H. Ten millimoles (1.80 grams)of caffeic acid was dissolved in 300 milliliters of 0.1 Normal aqueoussodium hydroxide (NaOH). The remaining procedure followed that ofExample 1. The yield of synthetic soil extract was 0.65 gram.

EXAMPLE 6 Preparation of a Synthetic Humic Acid fromTetrahydroxybenzoquinone

The chemical structure of the starting organic compound is shown inTable 2. Ten millimoles (1.72 grams) of tetrahydroxybenzoquinone wasdissolved in 300 milliliters of 0.1 Normal aqueous sodium hydroxide(NaOH). The remaining procedure followed that of Example 1. The yield ofsynthetic soil extract was 0.016 gram.

EXAMPLE 7 Preparation of a Synthetic Humic Acid from1,4-dihydroxybenzene (Hydroquinone)

The starting organic compound is shown in Table 1, and represented byR₁, R₄=—OH; and R₂, R₃, R₅, R₆=—H. Ten millimoles (1.10 grams) ofhydroquinone was dissolved in 300 milliliters of 0.1 Normal aqueoussodium hydroxide (NaOH). The remaining procedure followed that ofExample 1. The yield of synthetic soil extract was 0.16 gram.

EXAMPLE 8 Preparation of a Synthetic Humic Acid from3,4,5-trihydroxybenzenoic Acid (Gallic Acid)

The starting organic compound is shown in Table 1, and represented byR₁=—CH₂CO₂H; R₃, R₄, R₅=—OH; and R₂, R₆=—H. Ten millimoles (1.70 grams)of gallic acid was dissolved in 300 milliliters of 0.1 Normal aqueoussodium hydroxide (NaOH). The remaining procedure followed that ofExample 1. The yield of synthetic soil extract was 0.10 gram.

EXAMPLE 9 Preparation of a Synthetic Humic Acid from2,5-dihydroxyphenylacetic Acid (Homogentisic Acid)

The starting organic compound is shown in Table 1, and consisted ofR₁=—CH₂CO₂H; R₂, R₅—OH; and R₃, R₄, R₆=—H. Ten millimoles (1.68 grams)of homogentisic acid was dissolved in 300 milliliters of 0.1 Normalaqueous sodium hydroxide (NaOH). The remaining procedure followed thatof Example 1. The yield of synthetic soil extract was 0.20 gram.

The following Examples 10-13 are illustrative of the entire process ofthe preferred embodiments including step e), addition of salts. Additionof salts increases the activity of the synthetic humic acids produced bythe method. Examples 10-13 illustrate that the synthetic humic acidmaterials produced according to the chemical processes and separationand isolation procedures of the preferred embodiments exhibit thephysicochemical properties and attributes of typical naturally-occurringcommercially-available humic acids and other soil extracts. Examples10-13 also illustrate that the therapeutic indications of the synthetichumic acids produced according to the chemical processes and separationand isolation procedures of the preferred embodiments are those of soilextracts and humic acids in general, that is to say for viral-relateddisorders and diseases.

EXAMPLE 10 Preparation of Another Synthetic Humic Acid from2,5-dihydroxyphenylacetic Acid (Homogentisic Acid)

The starting organic compound is shown in Table 1, and represented byR₁=—CH₂CO₂H; R₂, R₅=—OH; and R₃, R₄, R₆=—H. Six millimoles (1 gram) ofhomogentisic acid was dissolved in 300 milliliters of 0.1 Normal aqueoussodium hydroxide (NaOH). The solution pH was adjusted to 8.5 with 6Normal HCl. One and one-half millimoles (0.32 gram) of sodium periodate(NaIO₄) and 0.12 gram of sodium sulfide nonahydrate (Na₂S.9H₂O; 0.5millimole) were added, and the solution was placed in a water-bath at50° C. overnight. One milligram of boric acid (H₃BO₃; 0.016 millimole),0.021 gram of ferrous sulfate heptahydrate (FeSO₄.7H₂O; 0.075millimole), and 0.006 gram of calcium sulfate dihydrate (CaSO₄.2H₂O;0.035 millimole) were added and the solution was stirred for 2 hours atroom temperature. Any precipitate was removed by centrifugation. Thesolution was dialyzed with a 1,000-dalton cut-off flow-throughopen-channel or screen membrane system (Pall Filtron: Ultrasette® 7Tangential Flow Device or Mini-Ultrasette® 7 Tangential Flow Device usedwith a Pall Filtron Ultralab® 7 Specialized Pump and Reservoir System)to a conductivity of 30 microsiemens or less against distilled water.The dialysis apparatus was then used to concentrate the solution toabout 200 milliliters. The solution can be saved at this point forfurther use as an aqueous solution; or it can be freeze-dried to apowder. (Fifty to two hundred milligrams of mannose or other suitablecarbohydrate can be added to the solution prior to freeze-drying toreduce static electricity effects associated with the freeze-driedpowder.) The yield of synthetic soil extract was 0.23 gram.

The HPLC trace of the synthetic soil extract obtained in this Example isillustrated in FIG. 1. Peaks 1-6 were produced by this example. Peak 5is under the shoulder of Peak 4 and is not overtly apparent. Amathematical first derivative of the detector signal versus time canmore clearly show Peak 5. FIG. 2 shows the HPLC trace of a typicalcommercially-available natural humic acid. Peak 6 in FIGS. 1 and 2 wasproduced by a column wash with 90-100% v/v methanol and also containssynthetic humic acid. It can be seen that with the exception of therelative amounts of material in Peaks 2, 4, and 6, the remainder of theHPLC traces in FIGS. 1 and 2 are essentially equivalent. Thus, thesynthetic procedure of the present invention produced a humic acidmaterial with physicochemical characteristics that are essentiallyequivalent to those of a commercially-available soil extract.

EXAMPLE 11 Preparation of Still Another Synthetic Humic Acid from2,5-dihydroxyphenylacetic Acid (Homogentisic Acid)

The starting organic compound is shown in Table 1, and represented byR₁=—CH₂CO₂H; R₂, R₅=—OH; and R₃, R₄, R₆=—H. Ten millimoles (1.68 grams)of homogentisic acid was dissolved in 300 milliliters of 0.1 Normalaqueous sodium hydroxide (NaOH). The remaining procedure followed thatof the preceding Example. The yield of synthetic soil extract was 0.47gram. The HPLC trace of the synthetic soil extract obtained in thisExample was identical to that described in Example 10 and illustrated inFIG. 1.

EXAMPLE 12 Preparation of Still Another Synthetic Humic Acid from2,5-Dihydroxyphenylacetic Acid (Homogentisic Acid)

The starting organic compound is shown in Table 1, and represented byR₁=—CH₂CO₂H; R₂, R₅=—OH; and R₃, R₄, R₆=—H. Ten millimoles (1.68 grams)of homogentisic acid was dissolved in 300 milliliters of 0.1 Normalaqueous sodium hydroxide (NaOH). The remaining procedure followed thatof the preceding Example. The yield of synthetic soil extract was 0.4gram. The HPLC trace of the synthetic soil extract obtained in thisExample was identical to that described in Example 10 and illustrated inFIG. 1.

EXAMPLE 13 Preparation of Another Synthetic Humic Acid from3,4-dihydroxycynnamic Acid (Cafeic Acid)

The starting organic compound is shown in Table 1, and represented byR₁=CHCHCO₂H; R₃, R₄=—OH; and R₂, R₅, R₆=—H. Ten millimoles (1.8 grams)of homogentisic acid was dissolved in 300 milliliters of 0.1 Normalaqueous sodium hydroxide (NaOH). The remaining procedure followed thatof the preceding Example. The yield of synthetic soil extract was 0.51gram.

EXAMPLE 14 Preparation of a Synthetic Humic Acid from1,3,4,5-tetrahydroxycyclohexane-carboxylic Acid3-(3,4-dihydroxycinnamate) (Chlorogenic Acid)

The starting organic compound is shown in Table 2. Ten millimoles (3.54grams) of chlorogenic acid was dissolved in 300 milliliters of 0.1Normal aqueous sodium hydroxide (NaOH). The remaining procedure followedthat of the preceding Example. The yield of synthetic soil extract was0.23 gram.

EXAMPLE 15 In vitro Toxicity of Synthetic Humic Acid Prepared Accordingto Examples 10, 11 and 12

Humic acid synthesized from homogentisic acid was prepared according tothe procedure of Examples 10, 11 and 12. The in vitro toxicity of thematerials was assessed as follows:

Five units of 450 milliliters each of whole human blood were collectedinto CP2D/AS-3 Leukotrap RC-PL systems. The blood was rested for 3 hoursat room temperature. Each sample was weighed, and then centrifuged at2820 revolutions per minute (2312 gravities) for 3 minutes, 44 seconds.The blood samples were then expressed through ATS-LPL filters intoplatelet storage bags. The filtration time was noted. The LR-PRP wascentrifuged at 3600 revolutions per minute (3768 gravities) for 7minutes. All but about 55 grams of platelet poor plasma was removed fromeach sample. The platelet concentrates were rested for 90 minutes atroom temperature, and were then weighed and placed in a plateletincubator. RCM1 filters were primed with AS-3 solution. The primary bagswere hung at a height of 60 inches above empty AS-3 bags, such thatfiltration occurred by gravity. The filtration time was noted, and theLRRCC systems were sealed off 3 inches below the RCM1 filters. Each RCM1filter together with 6 inches of tubing and the LR-RCC, including thedonor identification tube segment, were weighed. Samples were taken atthis point for post-filtration testing (LR-RCC).

At Day 1 sufficient synthetic humic acid was added to each plateletconcentrate so as to make its concentration 25 micrograms permilliliter. Treated platelet concentrates were then incubated in aplatelet incubator for 1 hour, following which samples of each plateletconcentrate were taken for testing. Subsequent samples were also takenon day 5 for further testing.

Table 3 shows the effect of the synthetic humic acid prepared asdescribed in Example 10 on the viability of platelet concentrates asmeasured according to the procedures of this Example. The results wereall nominal, that is, the synthetic humic acid had no effect on plateletviability (i.e., is nontoxic). The same results were obtained when theconcentration of humic acid was made 100 micrograms per milliliterinstead of 25 micrograms per milliliter. These results are particularlynoteworthy, as blood platelets are known to be sensitive to a variety ofchemical agents. It is for this reason that few safe anti-viraltreatments are available for blood platelets.

TABLE 3 pH at 22° C. pCO₂, mm Hg pO₂, mm Hg HCO3, mmol/L MPV, fl UnitNo. Day 1 Day 5 Day 1 Day 5 Day 1 Day 5 Day 1 Day 5 Day 1 Day 5 1 7.4667.394 19.3 12.8 33.5 44.4 16.8 9.5 7.0 6.6 2 7.321 7.215 21.6 14.3 9.922.2 13.8 7.3 6.7 6.3 3 7.320 7.276 24.4 16.6 10.3 21.3 15.6 9.7 6.7 6.54 7.368 7.308 20.7 14.3 13.4 22.2 14.6 8.9 6.5 6.3 5 7.457 7.454 20.113.8 23.7 29.0 17.1 11.6 7.7 7.4 Mean 7.386 7.329 21.2 14.4 18.2 27.815.6 9.4 6.9 6.6 Std. Dev. 0.071 0.095 2.0 1.4 10.2 9.8 1.4 1.5 0.5 0.6WBC Platelet Yield, ×10⁵ Yield, ×10¹⁰ Streaming % ESC % HSR Lactate,mmol/L Unit No. Day 1 Day 1 Day 5 Day 1 Day 5 Day 1 Day 5 Day 1 Day 5Day 1 Day 5 1 0.1 8.3 9.0 3 3 24.2 16.9 78.0 64.0 5.1 12.1 2 0.2 14.514.2 3 3 27.5 20.3 81.7 71.5 6.6 13.4 3 0.4 13.3 13.4 3 3 28.7 26.3 81.779.4 6.3 12.4 4 0.3 11.7 12.3 3 2 22.1 19.2 81.4 77.1 6.6 13.1 5 0.3 8.99.1 3 3 19.1 14.4 74.7 70.2 4.5 9.7 Mean 0.3 11.3 11.6 3.0 2.8 24.3 19.479.5 72.4 5.8 12.1 Std. Dev. 0.1 2.7 2.4 0.0 0.4 3.9 4.5 3.1 6.1 1.0 1.4

EXAMPLE 16 In vitro Toxicity of Synthetic Humic Acid Prepared Accordingto Examples 10-14

Humic acid synthesized from homogentisic acid was prepared according tothe procedure of Examples 10-12. Humic acid synthesized from caffeicacid was prepared according to the procedure of Example 13. Humic acidsynthesized from chlorogenic acid was prepared according to theprocedure of Example 14. Natural-product humic acid was prepared bydialysis with subsequent freeze-drying as described in Examples 1-14.The in vitro toxicity of the materials was assessed as follows:

Cytotoxicity was examined with six concentrations of each humatematerial, and one “no-drug” concentration. All materials were tested inAfrican green monkey kidney cells (CV-1; Diagnostic Hybrids, Inc.,Athens, Ga.) in triplicate. The cells were provided in flat dishescontaining multiple cell wells. The cells were cultured in the presenceof different concentrations of humate materials for 24-36 hours at35-37° C. in a CO₂-humidified incubator. The morphology of the culturedcells was examined visually to determine any cytotoxic effects. Noabnormal cell morphology was observed in cultures with “no drug” nor inany containing humate concentrations up to 500 micrograms permilliliter. Furthermore, no apparent CV-1 cell death (that is, celldetachment from the bottom of the wells) was observed at anyconcentration of any material tested. The results established that thematerials were not cytotoxic at concentrations up to at least 500micrograms per milliliter.

EXAMPLE 17 In vivo Toxicity of Synthetic Humic Acid Prepared Accordingto Examples 10-14

Humic acid synthesized from homogentisic acid was prepared according tothe procedure of Examples 10-12. Humic acid synthesized from caffeicacid was prepared according to the procedure of Example 13. Humic acidsynthesized from chlorogenic acid was prepared according to theprocedure of Example 14. Natural-product humic acid was prepared bydialysis with subsequent freeze-drying as described in Examples 1-14.

The in vivo acute intravenous systemic toxicity of the humate materialswas assessed as follows. Each humate material was dissolved separatelyin sterile, pyrogen-free 0.9% aqueous sodium chloride solvent to yieldsolutions of final concentrations of 1, 0.5 and 0.25 milligrams permilliliter. The test animals were viral antibody-free Swiss Webstermice, which weighed in the range of 17-23 grams at the time of testing.All test animals were quarantined and checked for signs of disease priorto testing. All test animals were group-housed five per cage in plasticcages with stainless steel suspended lids.

For each dose of each humate material, ten mice (five males and fivefemales) were administered the sample humate material intravenously inthe amount of 50 milliliters per kilogram body weight. Ten additionalmice were similarly administered 0.9% sodium chloride solution (thesolvent vehicle) as a zero control. This procedure resulted in humatedoses of 50 milligrams per kilogram body weight from the 1 milligram permilliliter solution, 25 milligrams per kilogram from the 0.5 milligramper milliliter solution, 12.5 milligrams per kilogram from the 0.25milligram per milliliter solution, and 0 milligrams per kilogram fromthe 0.9% sodium chloride (blank) solution.

Following injection, the mice were offered a balanced Teklad diet andwater ad libitum for the duration of the study. All mice were examinedfor viability for fouteen days. Zero time, day seven and day fourteenweights and toxic symptoms were recorded.

No mortalities were observed for any of the mice over the fourteen dayobservation period and, while some clinical findings were observed, theywere not indicative of toxicity.

EXAMPLE 18 Influenza A Cytoprotection Properties of Natural-Product andSynthetic Humic Acids Prepared According to Examples 10-14

Humic acid synthesized from homogentisic acid was prepared according tothe procedure of Examples 10-12. Humic acid synthesized from caffeicacid was prepared according to the procedure of Example 13. Humic acidsynthesized from chlorogenic acid was prepared according to theprocedure of Example 14. Natural-product humic acid was prepared bydialysis with subsequent freeze-drying as described in Examples 1-14.The anti-viral properties of the humate materials were assessedaccording to the following methods:

Influenza virus type A (Beijing/262/95) (H1N1) was obtained from theCenter for Disease Control (CDC). Influenza virus type A(Panama/2007/99) (H3N2) was also obtained from CDC. The test cell lineconsisted of Madin Darby canine kidney (MDCK) cells.

The first protocol used to determine the cytoprotection properties ofthe humate materials was as follows: each test was run in 96-wellflat-bottomed microplates. Four log₁₀ dilutions of each test compoundwere added to 3 cups containing the cell monolayer. Within 5 minutes thevirus was added and the plate sealed, incubated at 37° C., and the druginhibition of the viral cytopathic effect (CPE) assessed microscopicallywhen untreated infected controls had developed a 3 to 4+ CPE(approximately 72 to 120 hours). The data were expressed as 50%effective inhibitory concentration (CPE EC₅₀).

The second cytoprotection assay protocol, employed to validate the CPEresults, was as follows: in the neutral red dye uptake version of theantiviral test protocol described immediately above, the same 96-wellmicroplates were utilized after the CPE had been read. Neutral red dyewas then added to the medium: cells not damaged by virus take up agreater amount of dye, which can be read on a computerized micro plateautoreader (the method is described in detail by McManus: Appl. Environ.Microbiol. 1976, 31, 35-38). The date were expressed as 50% effectiveinhibitory concentration (NR EC₅₀).

The results of drug efficacy testing against influenza A viruses withthe above-described protocols are summarized in Table 4.

This Example demonstrates that the humate materials can protect cellsagainst infection by influenza A viruses.

TABLE 4 Influenza A (H1N1) Influenza A (H3N2) Compound¹ CPE EC₅₀ NR EC₉₉CPE EC₅₀ NR EC₉₉ Caffeic Acid Humate 1 0.6 <1 <1 Chlorogenic Acid 45 406 6.5 Humate Homogentisic Acid 3.7 3.2 4.5 3.2 Humate Natural-ProductHumate 2.5 2.5 <1 <1 ¹All concentrations in micrograms per milliliter.

EXAMPLE 19 Influenza B Cytoprotection Properties of Natural-Product andSynthetic Humic Acids Prepared According to Examples 10-14

Humic acid synthesized from homogentisic acid was prepared according tothe procedure of Examples 10-12. Humic acid synthesized from caffeicacid was prepared according to the procedure of Example 13. Humic acidsynthesized from chlorogenic acid was prepared according to theprocedure of Example 14. Natural-product humic acid was prepared bydialysis with subsequent freeze-drying as described in Examples 1-14.The anti-viral properties of the humate materials were assessedaccording to the following methods:

Influenza virus type B (Beijing/184/93) (H1N1) was obtained from theCenter for Disease Control (CDC). The test cell line consisted of MadinDarby canine kidney (MDCK) cells.

The two protocols used to determine the cytoprotection properties of thehumate materials were as described in Example 18.

The results of drug efficacy testing against influenza B viruses withthe above-described protocols are summarized in Table 5.

This Example demonstrates that the humate materials can protect cellsagainst infection by influenza B viruses.

TABLE 5 Influenza B Compound¹ CPE EC₅₀ NR EC₉₉ Caffeic Acid Humate <1 <1Chlorogenic Acid Humate 5.5 4.7 Homogentisic Acid Humate 3.2 3.2Natural-Product Humate <1 <1 ¹All concentrations in micrograms permilliliter.

The Examples described above establish relevant influenza antiviral dataand efficacy of the synthetic humate compounds. The scope of the studiesconforms with current requirements put forth by the FDA for pre-clinicalanalysis of new anti-virals.

Various modifications and alterations of this invention will be apparentto those skilled in the art without departing from the scope and spiritof this invention. It should be understood that the invention is notlimited to the embodiments disclosed therein, and that the claims shouldbe interpreted as broadly as the prior art allows.

What is claimed is:
 1. A method for inhibiting and/or treating influenzavirus infections in a mammal comprising administering to the mammal aneffective amount of a synthetic phenolic polymeric material, saidsynthetic phenolic material having been prepared by the following steps:a) dissolving in an aqueous solution at least one starting organiccompound comprising at least one hydroxyl group and at least onecarbonyl group or at least two hydroxyl groups on an aromatic structure;b) adjusting the pH of the aqueous solution resulting from step a) tobetween about 8 and 11; c) oxidizing the at least one starting organiccompound solution resulting from step b); d) polymerizing the oxidizedcompound from step c); e) adding at least one water soluble compound orsalt selected from the group consisting of boric acid, borate salts,alkaline earth salts, transition metal salts, alkaline sulfides,alkaline earth sulfides, or transition metal sulfides to the aqueoussolution resulting from step d); and f) removing molecules from thesolution resulting from step e below about 500 to about 10,000 daltons.2. The method according to claim 1, wherein the starting organiccompound is selected from the group consisting of a compound representedby the Formula I:

wherein R₁, R₂, R₃, R₄, R₅, and R₆ is a substituent selected from thegroup consisting of H, CH₃, CH₂CH₃, (CH₂)₂CH₃, CH(CH₃)₂, OH, OCH₃, CHO,CO₂H, CO₂CH₃, CH₂OH, CH₂OCH₃, CH₂CHO, CH₂CO₂H, CH₂CO₂CH₃, (CH₂)₂OH,(CH₂)₂OCH₃, (CH₂)₂CHO, (CH₂)₂CO₂H, (CH₂)₂CO₂CH₃, CH(CH₃)OH, CH(CH₃)OCH₃,CH(CH₃)CHO, CH(CH₃)CO₂H, CH(CH₃)CO₂CH₃, CH(CH₃)CH₂OH, CH(CH₃)CH₂OCH₃,CH(CH₃)CH₂CHO, CH(CH₃)CH₂CO₂H, CH(CH₃)CH₂CO₂CH₃, CH(OH)₂, CH(OH)OCH₃,CH(OH)CHO, CH(OH)CO₂H, CH(OH)CO₂CH₃, CH(OCH₃)OH, CH(OCH₃)₂, CH(OCH₃)CHO,CH(OCH₃)CO₂H, CH(OCH₃)CO₂CH₃, CH(OH)CH₂OH, CH(OH)CH₂OCH₃, CH(OH)CH₂CHO,CH(OH)CH₂CO₂H, CH(OH)CH₂CO₂CH₃, CH(OCH₃)CH₂OH, CH(OCH₃)CH₂OCH₃,CH(OCH₃)CH₂CHO, CH(OCH₃)CH₂CO₂H, CH(OCH₃)CH₂CO₂CH₃, (CH₂)₃OH,—(CH₂)₃OCH₃, (CH₂)₃CHO, (CH₂)₃CO₂H, (CH₂)₃CO₂CH₃, CHCHOH (cis or trans),CHCHOCH₃ (cis or trans), CHCHCHO (cis or trans), CHCHCO₂H (cis ortrans), CHCHCO₂CH₃ (cis or trans), CH₂CHCHOH (cis or trans), CH₂CHCHOCH₃(cis or trans), CH₂CHCHCHO (cis or trans), CH₂CHCHCO₂H (cis or trans),and CH₂CHCHCO₂CH₃ (cis or trans).
 3. The method according to claim 2,wherein the compound comprises at least one hydroxyl group and at leastone carboxylic acid group.
 4. The method according to claim 1, whereinthe starting organic compound is selected from the group consisting of


5. The method according to claim 1, wherein the aqueous solution in stepa) comprises sodium hydroxide.
 6. The method according to claim 1,wherein the temperature in step d) is between about 35° C. and 80° C. 7.The method according to claim 1, wherein the temperature in step f) isbetween about 20° C. and 80° C.
 8. The method according to claim 1,wherein the method of preparation of the synthetic phenolic polymericmaterial further comprises a step, following step g), of: h)concentrating the solution resulting from step g).
 9. The methodaccording to claim 8, wherein the method of preparation of the syntheticphenolic polymeric material further comprises a step, following step h),of: i) removing the water from the solution resulting from the step h).10. The method according to claim 1, wherein the influenza virusinfection is effected by a virus selected from the group consisting ofinfluenza A and influenza B.
 11. The method according to claim 1,wherein the mammal is a human.
 12. The method according to claim 1,wherein administering the effective amount of synthetic phenolicpolymeric material is performed systemically.
 13. The method accordingto claim 1, wherein administering the effective amount of syntheticphenolic polymeric material is performed topically.
 14. The methodaccording to claim 1, further comprising administering an additionalantiviral composition in combination with the effective amount of asynthetic phenolic polymeric material.
 15. The method according to claim14, wherein administering the effective amount of synthetic phenolicpolymeric material and the antiviral composition is performedsystemically.
 16. The method according to claim 14, whereinadministering the effective amount of synthetic phenolic polymericmaterial and the antiviral composition is performed topically.
 17. Amethod of inhibiting influenza viral attachment to host cells in amammal comprising administering to the mammal an effective amount of asynthetic phenolic polymeric material, said synthetic phenolic materialhaving been prepared by the following steps: a) dissolving in an aqueoussolution at least one starting organic compound comprising at least onehydroxyl group and at least one carbonyl group or at least two hydroxylgroups on an aromatic structure; b) adjusting the pH of the aqueoussolution resulting from step a) to between about 8 and 11; c) oxidizingthe at least one starting organic compound solution resulting from stepb); d) polymerizing the oxidized compound from step c); e) adding atleast one water soluble compound or salt selected from the groupconsisting of boric acid, borate salts, alkaline earth salts, transitionmetal salts, alkaline sulfides, alkaline earth sulfides, or transitionmetal sulfides to the aqueous solution resulting from step d); and f)removing molecules from the solution resulting from step e below about500 to about 10,000 daltons.
 18. The method according to claim 17,wherein the starting organic compound is selected from the groupconsisting of a compound represented by the formula I:

wherein R₁, R₂, R₃, R₄, R₅, and R₆ is a substituent selected from thegroup consisting of H, CH₃, CH₂CH₃, (CH₂)₂CH₃, CH(CH₃)₂, OH, OCH₃, CHO,CO₂H, CO₂CH₃, CH₂OH, CH₂OCH₃, CH₂CHO, CH₂CO₂H, CH₂CO₂CH₃, (CH₂)₂OH,(CH₂)₂OCH₃, (CH₂)₂CHO, (CH₂)₂CO₂H, (CH₂)₂CO₂CH₃, CH(CH₃)OH, CH(CH₃)OCH₃,CH(CH₃)CHO, CH(CH₃)CO₂H, CH(CH₃)CO₂CH₃, CH(CH₃)CH₂OH, CH(CH₃)CH₂OCH₃,CH(CH₃)CH₂CHO, CH(CH₃)CH₂CO₂H, CH(CH₃)CH₂CO₂CH₃, CH(OH)₂, CH(OH)OCH₃,CH(OH)CHO, CH(OH)CO₂H, CH(OH)CO₂CH₃, CH(OCH₃)OH, CH(OCH₃)₂, CH(OCH₃)CHO,CH(OCH₃)CO₂H, CH(OCH₃)CO₂CH₃, CH(OH)CH₂OH, CH(OH)CH₂OCH₃, CH(OH)CH₂CHO,CH(OH)CH₂CO₂H, CH(OH)CH₂CO₂CH₃, CH(OCH₃)CH₂OH, CH(OCH₃)CH₂OCH₃,CH(OCH₃)CH₂CHO, CH(OCH₃)CH₂CO₂H, CH(OCH₃)CH₂CO₂CH₃, (CH₂)₃OH,—(CH₂)₃OCH₃, (CH₂)₃CHO, (CH₂)₃CO₂H, (CH₂)₃CO₂CH₃, CHCHOH (cis or trans),CHCHOCH₃ (cis or trans), CHCHCHO (cis or trans), CHCHCO₂H (cis ortrans), CHCHCO₂CH₃ (cis or trans), CH₂CHCHOH (cis or trans), CH₂CHCHOCH₃(cis or trans), CH₂CHCHCHO (cis or trans), CH₂CHCHCO₂H (cis or trans),and CH₂CHCHCO₂CH₃ (cis or trans).
 19. The method according to claim 18,wherein the compound comprises at least one hydroxyl group and at leastone carboxylic acid group.
 20. The method according to claim 17, whereinthe starting organic compound is selected from the group consisting of


21. The method according to claim 17, wherein the aqueous solution instep a) comprises sodium hydroxide.
 22. The method according to claim17, wherein the temperature in step d) is between about 35° C. and 80°C.
 23. The method according to claim 17, wherein the temperature in stepf) is between about 20° C. and 80° C.
 24. The method according to claim17, wherein the method of preparation of the synthetic phenolicpolymeric material further comprises a step, following step g), of: h)concentrating the solution resulting from step g).
 25. The methodaccording to claim 24, wherein the method of preparation of thesynthetic phenolic polymeric material further comprises a step,following step h), of: i) removing the water from the solution resultingfrom the step h).
 26. The method according to claim 17, wherein theinfluenza virus infection is effected by a virus selected from the groupconsisting of influenza A and influenza B.
 27. The method according toclaim 17, wherein the mammal is a human.
 28. The method according toclaim 17, wherein administering the effective amount of syntheticphenolic polymeric material is performed systemically.
 29. The methodaccording to claim 17, wherein administering the effective amount ofsynthetic phenolic polymeric material is performed topically.
 30. Themethod according to claim 17, further comprising administering anadditional antiviral composition in combination with the effectiveamount of a synthetic phenolic polymeric material.
 31. The methodaccording to claim 30, wherein administering the effective amount ofsynthetic phenolic polymeric material and the antiviral composition isperformed systemically.
 32. The method according to claim 30, whereinadministering the effective amount of synthetic phenolic polymericmaterial and the antiviral composition is performed topically.
 33. Themethod according to claim 1, wherein the synthetic phenolic material hasbeen prepared by a method wherein step c) comprises adding an alkalineperiodate salt or alkaline-earth periodate salt to the aqueous solutionresulting from step b).
 34. The method according to claim 1, wherein thesynthetic phenolic material has been prepared by a method wherein stepd) comprises maintaining the temperature of the solution from step c)between about 20° C. and 100° C. for a period of at least about 30minutes.
 35. The method according to claim 1, wherein the syntheticphenolic material has been prepared by a method wherein by a methodfurther comprising allowing the aqueous solution from step e) to standwith or without stirring at about 20° C. to 100° C. for at least about 2hours after step e).
 36. The method according to claim 17, wherein thesynthetic phenolic material has been prepared by a method wherein stepc) comprises adding an alkaline periodate salt or alkaline-earthperiodate salt to the aqueous solution resulting from step b).
 37. Themethod according to claim 17, wherein the synthetic phenolic materialhas been prepared by a method wherein step d) comprises maintaining thetemperature of the solution from step c) between about 20° C. and 100°C. for a period of at least about 30 minutes.
 38. The method accordingto claim 17, wherein the synthetic phenolic material has been preparedby a method wherein by a method further comprising allowing the aqueoussolution from step e) to stand with or without stirring at about 20° C.to 100° C. for at least about 2 hours after step e).